Preparation of zein solutions directly from gluten



Patented May 29, 1945 PREPARATION (F ZEIN SGLUTIONS DIRECTLY FBQM @LUTEN Roy E. @oleman, Chicago, Ill., assignor to Time, Incorporated, a corporation oil New York No Drawing. @riglnal application-November 30,

1942, Serial No.

467,402, new Patent No.

2,352,604, dated July 4, 1944, Divided and this application October 25, 1943, Serial No. 507,600

@laims.

This invention relates to methods of preparing commercially usable solutions and coating compositions of prolamines or prolamine-containing proteins directly from glutens, either corn, wheat. barley, etc. It is particularly directed to such methods which will produce solutions and coating compositions of the alcohol-soluble portion of corn gluten, of controlled solution and gelling characteristics.

This application is a division of my application, Serial No. 467,402, filed November 30, 1942, now Patent No. 2,352,604, dated July 4, 1944.

Heretofore, in the preparation of zein solutions, and coating compositions, including the substantially non-aqueous zein solutions and coating compositions of controlled solution and gelling characteristics as described in my prior Patent No. 2,185,110, granted December 26, 1939, for example, the zein is initially isolated from corn gluten and then dissolved in suitable solvents to form the desired solutions. To isolate zein from corn gluten as a commercially usable powder is a laborious and expensive process, involving steps of precipitation, settling, filtering, washing and drying. The solubility of the zein in'zein solvents and the solution characteristics such as stability and gelling tendencies are largely dependent upon the care exercised in carryingout the steps of the process.

In accordance with the present invention I prepare solutions of zein or zein-containing proteins directly from corn gluten under conditions that e will readily produce a wide variety of zein products in so far as properties and solubility characteristics are concerned, and thereby avoid the expense and objections incident to the preliminary isolation or zein in powder form. In carrying out the present invention, corn gluten, for example, in granular or powdered form, is treated with a suitable relatively low boiling point zein protein solvent or mixture of zein protein solvents, without the establishment of special conditions of hydrogen ion concentration either in the solvent or in the gluten, to extract the zein from thetemperatures, say from about 120. to about 170 increased. 1

1''.. preferably between about 140 to about 150.

E, for a short period 0! time generally not exceeding about three hours. It is preferred that the extraction time be not longer than about onehalf to about one hour since within this time and at the temperature stated, the zein in the gluten is extracted and-is not deleteriously affected.

The extract solution obtained as above de-.

by filtration, pressing, centrifugal separation or in any other desirable manner, mixed with a relatively high boiling point zein base solvent to be hereinafter described and distilled or otherwise treated to remove all or some 0! the rela-' tively low boiling point solvent, as desired. The

resulting residue is a solution of zein or zein-containing protein in the zein base solvent alone withall oi therelatively low boiling point solvent removed, or in the zein base solvent and that portion of the relatively low boiling point solvent which has not been removed. The residual gluten,

which still contains some protein, may be freed from retained solvent by washing and recovered for further useas a cattle food or for other purposes.

The resulting zein or zein-containing solutions in accordance with the present invention, over a wide range of concentrations, are stable on standing at temperatures of 80 F. and below and do not separate even when cooled to temperatures of 50 to 70 F. and somewhat below. When cooled even to temperatures as low as 10 above zero or to zero, solutions prepared in accordance with the present invention may separate and become solid: however, on heating again to temperatures of from about to 70 a re-solution is efiect ed, either without stirring or with-slight stirring, and the solutions return to their normal form at the re-heatingiemperatures. In general, these solutions are substantially non-gelling and have but a slight tendency to increase in viscosity; that is, to thicken or become heavier in body. In many instances they exhibit practically no tendency to thicken or to gel over practical periodsof time in the order of about 3 to 6 months and even up to one year and longer. These solutions remain I stable and reasonably constant in their characteristics during the periods necessary for transportation and distribution, either in bulk or in packages, or for storage for reasonable periods or time, and hence their field of applicability -is greatly The protein content of gluten varies widely, depending largely upon the source of the gluten and the method of making it or obtaining it as a byproduct. In the case of corn gluten now on the market, the protein content varies from about 40% to 70%." or this protein content, about to is aqueous-alcohol soluble and is extractable as zein or zein-containing proteins. The amount of zein or zein-containing proteins extracted in accordance with the methods of'the present invention is largely influenced by theparticular scribed is now separated from the residual gluten as solvent or solvent mixture used to eflect extracvents concentrated alcohol solvents. I tracting solvents may also be one or more oi. the

2 ear-moi tion as well as by the amounts thereof. For most purposes, about 2% parts by weight solvent to about 1 part by weight oi gluten is sufllcient to remove substantially all of the extractable sein protein. More or less solvent may be used, as desired, as is evident iromthespeciilc illustrative examples herein.

then 158 lbs. of base solvent will be required for no mately 60% total protein, the extract solutions each 100 lbs. of gluten, providing the estimated amount 0! soluble protein is extracted. It is evident that the protein content oi. the extract solution may be determined prior to the admixture oi base solvent with the extract solution, thereby making possible the determination of the precise. amount of base solvent to be added to the extract solution to obtain the desired concentration of zein protein in the final solution.

The sein' protein extracting solvents in accordance with the present invention maybe,- suitably, a single solvent or a solvent mixture which is a solvent for zein or other prolamine and which has a boiling point below about 125' C. and, preterably, or about 100 C. and below. Thus, the extracting solvent may be one or more of the wellknown alcoholic sein solvents such as an rous methanol, or constant' boiling oint mixtures- (aseotropes) of aliphatic alcohols such as 95% ethyl alcohol and 81% isopropyl alcohol. For

oicalsintl'iebascsolventsisrcquiredtosecure convenience herein I term these alcoholic sol- The exconcentrated alcohol solvents containing additional amounts of water, say from 40% to 50% uents o! the mixture are non-solvents ior sein proteins. Thus, they may be aseotroplc mixtures of water on the one'hand and dioxan or butyl alcohol on the other. Dloxan iorms with water a constant boiling point mixture containing 50% dioxan boiling at 86.9 0.; and butyl alcohol forms with water a constant boiling point mixture containing 63% butyl alcohol boiling at 92' C.

a In general, extracting solvents consisting 0! or including the aqueous alcohol mixtures are capable oi extracting larger amounts of s ein pro tein from gluten than are the extracting solvents consisting of or including a concentrated alcohol solvent. It has been noted that for practical extraction procedures, using either a concentrated alcohol solvent alone or an aqueous alcohol mixture alone, approximately 2% parts by weight of solvent to 1 or gluten may be used. At this concentration, with gluten containing approxigs sein protein content of from about 12% to about 15% and even up to In addition, the aqueous alcohol mixture solvents are capable of extracting a substantially larser proportion of a desirable type of zein present in the protein of so the gluten than are the concentrated alcohol solvents.

4 The base solvents are relatively high boiling compounds or mixtures of compounds having a boiling point above about 125 (2., preferably. above about 150 C. and having a suitable balance between the hydrocarbon constituents or radicals present in the solvent or solvent mixtures and the polar constituents'or radicals present therein. A certain proportion of hydroxyl redand maintain solvent action. Other polar radicals such as O-(ether oxygen), Cl,- -NH:

and COOK appear to aid or cooperate with 'the OHinsecuringthenecessaryconditionsoi in anhydrous'methanol orin excess-0i that pres-- P y in the se s lv nts in whi h erm I ent in the aseotropes 05% alcoholand 01% tsopropyl alcohol. Greater or lesser amounts otwaterthanasstatedmaybeused; hcwever,bestresuits are obtained with the use otadded amounts of water within the percentagerange stated. For

aqueousalcohol V Anotherclassoiextractingsolventsinaccordconvenience herein I term these alcoholic. solvents ance with the present invention comprises mix-.

l gsolventslalsdincludemixturesotfi'k alco-' hol and non-solvents as disclosed in my prior Patent No. 2,185,123, granted December 25,1939, I andnon-aqucous aseotropic mixtures of anhy- 'drous methyl alcohol with hexane, cyclohexane.

trlchlcre thyienc, acetone, methyl acetate, sums acetate, metlwl ethyl ketone, etc., because one comtituentoithemixturam'ethflalcohohisa' sein solvent.

ln-additionto the foregoing, immanentone constituent of an extracting solvent .miximeisaseinsolvenhtheextractngsolventsmay be suitable aseotropic which are sein protein-solvents wherein the individual ccnst itinclude mixturesoi base solvents). for solvent action.

The proportion of polar radicals to hydrocarbon or non-polarradicals which I have toundto ll be desirable lieswithin a range which extends between the limits of absolute methanol on the onev hand, having 53% hydroxyl and 47% hydrocarbon radicals. and absolute ethanol on the other hand having 37% hydl'bl yl and 63% bylI-drocarbon radicals. The bensyl radical acts as it it were intermediate the methyl and ethyl rad- I icals. 'Ihis'rangeisapproximateandmayvary somewhat depending on the solvent or solvent mixtureused. Thelimitsoithisrangeareinso dicative of solvent properties and provide a close and adequate guide to enable the operator. by .simpleexperimentatiomtodetcrminetherequimd 'balanceorpolarandhydrocarbonradicalsinthe .bases olventormixtuteofbasesolvcntsinacoordes ance with the present invention.

Itmaybestated thatthcbasesolventsinaccordance'withthe pre'aentinvention may beany oneoithe solvent ormixtnre of solvents described in my prior Patent No. 2,155,110 whichhastherequiredbalanceoipolarandnon-polarradicals, and a boiling point above about 6., preferably, above about .C. as pointed out above. 'l or a more canplete description 0! the kindsoisolventswhichmaybeusedasbasesoll5 vents in accordance with the present invention 'conditionis particularlypronounced when the than is disclosed herein, reference isinade to the description of thse so nts inf myfiprior Patent No. 2,185,110.

1 Examples of base solvents n accordance with the present finventionjare' "the glycols such diethylene "glycol, propylene glycolfi'triethyleiiej lycol and mixtures thereof; including'mixtures which contain ethylene swear;placements; hol; close'd chain cyclic alcohols such-benzyl alcohol, cyclohexanol, furiuryT-alcohol, tetrah'y-j drofuriuryl' alcohol and mix ures ofclo'sedch'ain aicohols;"glycol e'thers' such as'ethylene glycol, mono-methyl ether, ditliylenfe glycol j monomethyl ether, ethylene g1ycq1 mona+emy1 ether, diethylene glycol monoethyl 'andniixtures of sly col others; mixtures of two oi-more ?oi'- the foregoing enumerated base solvents; mixtures oifone or more of the foregoing'bas'e solvents with glycerine, or other solvents or i mixtures" of I solvents, providing the mixture hasytne required polar w non-polar radical balance andtherequired; relatively high boiling point. i It is Qtobeunder'stood that the foregoing specificallymentiiined solvents or mixtures of solvents are' not "intended to" be inclusive of all the base*'solvents which maybe used in accordance with the present invention since they are merely *s'et forth for illustrative purposes. j r

Zein or zein-containing extract solutions containing any one or moreofthefore-going base solvents, in accordance with the present invention may be boiled, distilled or otherwise treated to remove part or all of? the." diliient. It is evident that practically all of the extracting solvent. including any water carried from i tlie gluten to the'e'xtract solution, may; be removed, thereby obtaining solutions or zein or: zein-containing proteins, in .the base solvent which, for all practical purposes, are substantially anhydrous. Such solutions vary widely in character, depending upon the choice of solvents or solvent mixtures used, as is to be expected, and some of them may undergo changes (apparent insolubility, for example), during 'the solvent' and water removal whicheflect the solvent characteristics of the zein and zein-containing proteins in the Esgsolvent utilized. These changes, where they ke place, are not of .a permanent character and it is possible to effect a re-solution of the zein or zein-containing'proteins inthe base sol-- vents by the addition of a small amount of water, sayin the order of from about 1 to 3% byweight, or an equivalent amount of other highly hydroxylated compound, such as methyl or ethyl alcohol or a mixture of such compounds.

In the preparation of-usable solutions having a wide held of applicability, I'prefer that the removal of the relatively lowboiling point solvent and water be controlled soas to leave in the final solutions at least about 1 to 3% by weight of water or an equivalent amount of other highly hydroxylated compound. preferably. water. Where this is not possible, water may be added to the final solution, in the amounts stated.

While water is, in general, not an essential con-.

character of the extraction solventsused.

highly aqueous alcohols or the hish'water azeotropes'such as dioxan-water or 'n-butyl alcoholwater mixtures are used assolvents. -Not only dogthese' solvents eflect the extraction of a larger i amount of the zein-containing protein than do the'more concentrated alcohols, but they also produce by their extraction atype of zein protein' which generally produces solutionshavin I afhigh viscosity and pronounced colloidal consistency. On the other hand the zein protein produced with the concentrated alcohols'and'less aqueous diluents tends to produce solutions having a lower viscosity and a lesser colloidal consistency. It is thus evident thatvarious types of zein protein having substantially difierent properties can be produced by proper s'election and composition of the extracting solvents.

From the foregoing it is manifest that the at ture and properties of the finished solution will be determined by the amount-of'zein protein present in the solution, the type or character of the zein protein extracted by the particular extracting solvent used and by the nature of the base solvent. It is apparent that with the extracting solvent and base solvent combinations possible, awide variety of solutions for manyuses can be produced by the method of the present invention.

Within the range of the many possible combinations and the method in accordance with the present invention, solutions may be produced.

which require further adjustment or modification in order to secure improved solution or desired'stability and controlled gelling properties. Such adjustments or-modifications canbe readily effected bythe use of rosin, fatty acids, compatible amines 'such as 'monoethanolamine, triethanolamine,=etc.,' amine soaps of'rosin or'fatty acids, or by the various methods and materials for producing stable, controlledgelling, zein'solutions described in my issud flatents Nos. 2,185,- 122, '-2,246,'I79, and 2,298,548 The; addition of suitable materials for e'il'ec'tin esiredmodiflcation-or adjustment may beriidtle the extract ples and elsewhere throughoutz 'the specification,

the term."parts" indicates ipartsfiby weight.

Example 1.400 parts ot- .91 isopropyl alcohol were added to 165' parts'foi'zgpowdered gluten in a vessel. 'Ihe 'mixture was,;,heated to about 180 1''. and held at-about that;='temperature for about 55 hour with stirringsjtczextract the aeincontaining proteins from the gluten. The mixture was then forced througha'fllter to separate stirring until cessation oiyisible boiling. 'rne the zein-containing extract. To 100 parts; of the extract 40 parts of propyleneglycol were added and the mixture was slowly heated to about 250 1". and held at about that temperature with residue was a solution of zein-containing proteins principally in propylene glycol.

' m the following examples the manipulative ,were the same, as set forth in Example 1 exoeptthat the extractions were carried gout at I140} v 1''. and the proportions varied .In each instance the point ible boiling.

mixtureo! extract andrelatively hlahbomm solvent was heated until cessation of visl'mple 2.--75 parts of powdered gluten were extracted with .187 parts of a solvent mixture consisting of 70% of denatured.95% ethyl alcohol (Bynasol), and 30% of water. 29%parts of diethylene glycol were added to 50 parts of theextract. TheprocedurewasasinExample 1 and the resulting solution was asolution off transparent coatings even when retaining some solvent. The coatings formed by these rein-containing solutions and coating compositions are very strong and have adherent properties. 5 In these solutions and coating compositions thedllm forming properties thereof can be built up to produce films of great toughness. flexibility, hardness and-gloss. Under normal temperature canrein-containing proteins principally in diethyllution of min-containing proteins principally in diethylene glycol monoethyl ether.

Emmple 5.30 parts of powdered gluten were extracted with 75 parts of a solvent mixture consisting of 80% o! dioxan and of water. parts of ethylene glycol monoethyl ether were added to 4'7 parts of the extract. The procedure was as in Example 1 and the resulting solution was a solution of rein-containing proteins principally in ethylene glycol monoethyl ether.

Example 6.-30 parts of powdered gluten were extracted with 75 parts of a solvent mixture consisting 'of 83% of n-butyl alcohol and 37% of water. 19.8 parts of benzyl alcohol were added to 44 parts of the extract. The procedure was as in Example 1 and the resulting solution was a solution of -vein-containing proteins principally in benzyl alcohol;

Example 7.-20 parts of powderedgluten were extracted with 50 parts of a. solvent mixture consisting of 60% of 91% isopropyl alcohol and-40% of water.- 17% parts of diethylene glycol were added to 85 parts of the extract. The procedure was as in Example 1 and the resulting solution was a solution of rein-containing proteinsprincipally in diethylene glycol.

At the cessation of boiling as referred to in.

the examples some proportion of the lower boiling constituents of. the diluent originally used remains in the final solution. The proportion thereof so remaining depends upon the final temperature and the boiling point curve of the mixture. Considerable proportions of the diluent or ofits constituents, including water, may remain without causing gelling of the final solution in reasonable periods of time. This is particularly true where the'glycols are used. In this respect the final solutions show dlflerences from solutions of commercial sein containing large amounts of water as in the prior art, apparently by reason of some changes in the character or structure of the rein-containing proteins of the corn gluten taking place in the process of the present invention, the nature of which changes is not as yet understood. a

changes in materials, proportions and method details will be apparent from the foregoing.

In the claims the expression "an elevated temperature in the order of about 120' I". to about 170 F. "is not to be construed as a precise critical range. but, rather, as a range of temperatures indicative of the elevated temperature at which the extractions may be carried out.

I claim:

1. The method of preparing directly from gluten a solution of min-containing proteins in a solvent or mixture of solvents comprising as an essential solvent constituent for the zeincontaining proteins, a solvent having a boiling point above about 125' C'. and having a proportion of polar to non-polar radicals which lies within a range extending betweentlie limits of the proportion of polar to non-polar radicals of methanol and the proportion of polar to nonpolar radicals of ethanol, which comprises con-- tacting gluten with an aqueous zein protein solvent having a boiling point below about 125 c. at an elevated temperature in the order of about 120 F. to about 170' P. to extract min-containin: proteins from the gluten, separating the extract solution from the residual gluten, mixing the aforesaid solvent having a boiling point above about 125 C. with said extract sohition and removing at le t part of said relatively low boiling-point solven from the solution.

In general and influencedby the type and 'amount of rein protein present,.the solutions tough. flexible, non-tacky, hard and generally 2. The method of preparing directly from gluten a solution of min-containing proteins in a solvent or mixture of solvents comprising as an essential solvent constituent for the sein-containing proteins, a solvent having a boiling point above about 150 C. and having a proportion of polar to non-polar radicals which lies within a range extending between the limits of the proportion of polar to non-polar radicals of methanol and the proportion of polar to non-polar radicals of ethanol, which comprises contacting gluten with an aqueous nein protein solvent having a boiling point below about C. at an .elevated temperature in the order of about 120' 1". to about the. residual gluten. mixing a rein-containing protein solvent comprising the aforesaid solvent having a boiling point above about C. .with said extract solution and removing substantially all of said relatively low boiling point solventfrom the solution.

a 3. The method 'of preparing directly from gluten a solution of rein-containing proteins in a solvent or mixture oi solvents comprising as an essential solvent constituent for the zein-containing proteins, a solvent having a boiling point above about 150 C. and having a proportion 'of polar to non-polar radicals which lies within a range extending between the limits of the proportion of polar to non-polar radicals of methanol and the proportion of polar to non-polar radicals of ethanol, which comprises contacting gluten with an aqueous zein protein solvent having a boiling point below about 125 C. at an elevated temperature in the order of about 120 F. to about 170 F. to extract zein-containing proteins from the gluten, separating the extract solution from the residual gluten, mixing a zeincontaining protein solvent comprising the aforesaid solvent having a boiling point above about 150 C. with said extract solution and removing at least part of said relatively low boiling point A solvent from the solution.

4. The method of preparing directly from gluten a solution of zein-containing proteins in a solvent or mixture of solvents comprising as an essential solvent constitutent for the zein-con-- taining proteins, a solvent having a boiling point 1 above about 150 C. and having a proportion of polar to non-polar radicals which lies within a range extending between the limits of the proportion of polar to non-polar radicals of methanol and the proportion of polar to non-polar radicals of ethanol, which comprises contacting gluten with an aqueous zein protein solvent having a boiling point below about 100 C. at an elevated temperature in the order of about 120 F. to about 170 F. to extract zein-containing proteins from the gluten, separating the extract solution from the residual gluten, mixing a zeincontaining protein solvent comprising the aforesaid-solvent having a boilingpoint above about 150 C. with said extract solution and removing at least part of said relatively low boiling point solvent from the solution.

5. The method or preparing directly irom gluten a solution of zein-containing proteins in a solvent or mixture 01' solvents comprising a glycol as an essential solvent constituent for the zeincontaining proteins, which comprises contacting gluten with an aqueous zein protein solvent having a boiling point below about 125 C. at an elevated temperature in the order of about 120 F. to about 170 F. to extract zein-containing proteins from the gluten, separating the extract solution from the residual gluten, mixing a zeincontaining protein solvent comprising a glycol with said extract solution and removing at least part of said relatively low boiling point solvent from the solution.

6. The method of preparing directly from gluten 5 solution or zein-containing proteins in a solvent or mixture or solvents comprising a glycol.

as an essential solvent constituent for the zeincontaining proteins, which comprises contacting cluten with an aqueous zein protein solvent having a boiling point below about 125 C. at

an elevated temperature in the order or about 120' l. to about 170 F. to extract coin-containproteins rrom the gluten, said zein protein solvent I comprising a mixture or a zein protein solvent and a non-solvent forzein, separating tbe'extractsolution from the reeidualgluten, mixing 0. Iaincontsining protein solvent comprising a glycolwith said extract solution and removin at least part of said relatively low boiling point solvent from the solution. i

7. The method of preparing directly from. glu ten a solution or zein-containing proteins in a solvent or mixture of solvents comprising a glycol as an essential solvent constituent for the zeincontaining proteins, which comprises contacting gluten with an aqueous zein protein solvent having a boiling point below about 125 C. at an elevated temperature in the order of about 120 F. to about 170 F. to extract rein-containing proteins from the gluten, said zein protein solvent comprising an azeotropic mixture of an alcohol having from two to three carbon atoms in the molecule and water, and a non-solvent forzein, separating the extract'solution from the residual gluten, mixing a zein-containing protein solvent comprising a glycol with said extract solution and removing at least part of said relatively low boiling point solvent from the solution.

8. The method of preparing directly from gluten a solution'of zein-containing proteins in a solvent or mixture of solvents comprising propylene glycol as an essential solvent constituent for the zein-containing proteins, which comprises contacting gluten with a relatively low boiling point zein protein solvent comprising 95% ethyl ene glycol monoethyl ether as an essential solvent constituent for the zein-containing proteins,

which comprises contacting gluten with a relatively low boiling point zein protein solvent comprising 95% ethyl alcohol and water at an ele-- vatedtemperature in the order of about 120 F. to about 170 F. to extract zein-containing proteins from the gluten, separating the extract solution from the residual gluten, mixing a zeim' containing protein solvent comprising diethylene glycol monoethyl ether with saidextract solution and removing at least part of said relatively low boiling point solvent from the solution.

10. The method of preparing directly from glu ten 9. solution of min-containing proteins in a solvent or mixture of solvents comprising diethylene glycol as an essential solvent constituent for the zein-containing proteins, which comprises contacting gluten with a relatively low boiling p int zein protein solvent comprising isopropyl alcohol and water at an elevated temperature in theorder or about F. to about F. toextract zein-containing proteins from the gluseparating the extract solution from the residual gluten, mixing a rein-containing protein solvent comprising diethylene glycol with said extract solution and removing at least part of said relatively low boiling point solvent from the solution.

BOY I. COLEMAN. 

